WF14573 or its salt, production thereof and use thereof

ABSTRACT

The present invention provides a new antimicrobial compound WF14573, and a process for producing WF14573 by (a) culturing a WF14573A and/or B-producing microorganism in a nutrient medium and recovering WF14573A and/or B from the resultant cultured broth or (b) deacylating WF14573A and/or B in the presence of a microbial substance which is capable of deacylating WF14573A and/or B. Also provided are an antimicrobial agent comprising WF14573 and carrier(s), a pharmaceutical composition comprising an effective amount of WF14573 and pharmaceutically acceptable carrier(s), a method for killing microorganisms by applying WF14573 to the microorganisms, and use of WF14573 for the treatment of infectious diseases caused by pathogenic microorganisms.

TECHNICAL FIELD

This invention relates to a new antimicrobial compound, WF14573 or itssalt. More particularly, it relates to a new antimicrobial compound,WF14573 or its salt which has an antimicrobial activity againstpathogenic microorganisms, especially pathogenic fungi, a process forthe preparation thereof and a pharmaceutical composition comprising thesame.

DISCLOSURE OF INVENTION

The new compound, WF14573 is represented by the following formula:

wherein R¹ is hydrogen or methyl and R² is hydrogen or palmitoyl.

In this specification, the following designations of the specificcompound are conveniently used.

Compound name R¹ R² WF14573 A —H —CO(CH₂)₁₄CH₃ WF14573 B —CH₃—CO(CH₂)₁₄CH₃ Deacyl WF14573 A —H —H Deacyl WF14573 B —CH₃ —H

The new compound, WF14573A has the following physico-chemicalproperties:

a) Molecular weight: ESI-MS(negative)m/z 1143 (M−H); b) Elementalanalysis: C 48.39; H 7.15; N 8.95; c) Melting point: 230-240° C. (dec.);d) Optical rotation: [α]_(D) ²³=−12 (c 0.5, methanol); e) UV spectrum: λmax(ε)=276 nm (methanol); f) IR spectrum: ν max(KBr)=3360, 2920, 2830,1670, 1630, 1540, 1440, 1270, 1240, 1050 cm⁻¹; g) ¹H-NMR spectrum: (500MHz, CD₃OD) δ (ppm): 7.18 (1H, d, J=8 Hz), 6.80 (1H, d, J=2 Hz), 6.67(1H, dd, J=8, 2 Hz), 5.30 (1H, d, J=3 Hz), 5.10 (1H, d, J=4 Hz), 4.98(1H, d, J=3 Hz), 4.64 (1H, m), 4.58˜4.52 (3H, m), 4.47˜4.43 (2H, m),4.40˜4.33(2H, m), 4.27 (1H, d, J=3 Hz), 4.02˜3.95 (3H, m), 3.82˜3.76(2H, m), 2.70 (1H, m), 2.63˜2.55 (2H, m), 2.48˜2.40 (2H, m), 2.30˜2.17(3H, m), 2.05˜1.93(4H, 1.57 (2H, m), 1.35˜1.23 (24H, m), 1.15 (3H, d,J=6 Hz), 0.89 (3H, t, J=7 Hz). h) ¹³C-NMR spectrum: (125 MHz, CD₃OD) δ(ppm): 176.8 (s), 175.9 (s), 174.4 (s), 174.1 (s), 172.9 (s), 172.7 (s),172.5 (s), 169.3 (s), 150.3 (s), 140.1 (s), 137.6 (s), 124.0 (d), 121.9(d), 119.4 (d), 74.2 (d), 74.0 (d), 73.9 (d), 71.3 (d), 70.7 (d), 70.5(d), 69.8 (d), 68.3(d) 62.4 (d), 58.5 (d), 58.0 (d), 57.2 (t), 55.4 (d),51.2 (d), 47.0 (t), 41.1 (t), 39.7 (t), 38.9 (t), 36.7 (t), 35.0 (t),34.6 (t), 33.1(t), 30.80 (t×5), 30.76 (t), 30.7 (t), 30.6 (t), 30.5 (t),30.3 (t), 27.0 (t). 23.7 (t), 19.5 (q), 14.4 (q). i) Solubility;Soluble: water, methanol. dimethylsulfoxide; Insoluble: n-hexane,chloroform; j) Thin layer chromatography: Carrier: Silica gel 60 F254(Merck); Solvent: 1-butanol: acetic acid: water=4:1:2; Rf=0.39.

The new compound, WF14573B (as its sodium salt), has the followingphysico-chemical properties:

a) Molecular weight: ESI-MS(negative)m/z 1157 (M−H); b)Elementalanalysis: C 48.79; H 7.34; N 8.96; S 2.80; Na 1.69; c) Melting point:220-225° C. (dec.); d)Optical rotation: [α]_(D) ²³=15° (c 0.9,methanol); e) UV spectrum: λ max(ε)=276 nm (methanol); f) IR spectrum: νmax(KBr)=3360, 2940, 2830, 1670, 1630, 1530, 1440, 1270, 1240, 1050cm⁻¹; g) ¹H-NMR spectrum: (500 MHz, CD₃OD) δ (ppm): 7.18 (1H, d, J=8Hz), 6.80 (1H, d, J=2 Hz), 6.67 (1H, dd, J=8, 2 Hz), 5.29 (1H, d, J=3Hz), 5.07 (1H, d, J=4 Hz), 4.99 (1H, d, J=4 Hz), 4.64 (1H, m), 4.58˜4.51(3H, m), 4.48˜4.36 (4H, m), 4.16 (1H, m), 4.07 (1H, m), 4.02˜3.97 (2H,m), 3.79 (1H, br. d, J=11 Hz), 3.37 (1H, m), 2.69 (1H, m), 2.60˜2.40(5H, m), 2.22 (2H, m), 2.05˜1.98 (3H, m), 1.57 (2H, m), 1.35˜1.24 (24H,m), 1.15 (3H, d, J=6 Hz),1.06 (3H, d, J=7 Hz), 0.89 (3H, t, J=7 Hz). h)¹³C-NMR spectrum: (125 MHz, CD₃OD) δ (ppm): 176.7 (s), 176.0 (s), 174.3(s), 174.1 (s), 172.8 (s), 172.5 (s), 172.5 (s), 169.3 (s), 150.3 (s),140.1 (s), 137.6 (s), 124.0 (d), 122.0 (d), 119.4 (d), 75.9 (d), 74.0(d), 73.9 (d), 71.3 (d), 70.7 (d), 70.5 (d), 70.2 (d), 68.2 (d), 62.4(d), 58.6 (d), 58.0 (d), 57.2 (t), 55.5 (d), 52.9 (t), 51.3 (d), 41.1(t), 39.8 (t), 39.1 (d), 38.9 (t), 36.7 (t), 34.9 (t), 33.0 (t), 30.8(t×5), 30.7 (t), 30.7 (t), 30.5 (t), 30.4 (t), 30.3 (t), 27.0 (t), 23.7(t), 19.5 (q), 14.4 (q), 11.1 (q). i) Solubility; Soluble: water,methanol, dimethylsulfoxide; Insoluble: n-hexane, chloroform; j) Thinlayer chromatography: Carrier: Silica gel 60 F254 (Merck); Solvent:1-butanol: acetic acid: water=4:1:2; Rf=0.45.

In the result of the above physicochemical properties and extensiveinvestigations, the chemical structures of WF14573A and B are identifiedas mentioned above, respectively.

BEST MODE FOR CARRYING OUT OF THE INVENTION

According to this invention, the compound, WF14573A and B can beprepared by culturing a WF14573A and/or B-producing strain belonging tothe genus Coleophoma in a nutrient medium.

Particulars of microorganisms used for the production of WF14573A and Band production thereof will be explained in the followings.

Microorganism

The microorganism which can be used for the production of WF14573A and Bis a WF14573A and/or B-producing strain belonging to the genusColeophoma, among which Coleophoma empetri No.14573 was newly isolatedfrom a decayed leaf sample, collected at Mitsushima-cho, Kamiagata-gun,Nagasaki-ken, Japan.

Lyophilized samples of the newly isolated microorganism, Coleophomaempetri No. 14573 was deposited with an International DepositoryAuthority on the Budapest Treaty, National Institute of Bioscience andHuman-Technology, 1-3. Higashi 1-chome, Tsukuba-shi, Ibaraki-ken, 305,Japan under the deposit number FERM BP-6252 on Feb. 12, 1998.

It is to be understood that the production of the new compound, WF14573Aand/or B is not limited to the use of the particular organism describedherein, which is given for illustrative purpose only. This inventionalso includes the use of any mutants which are capable of producing theWF14573A and/or B including natural mutants as well as artificialmutants which can be produced from the described organism byconventional means, such as genetic engineering, X-ray, ultravioletradiation, treatment with N-methyl-N′-nitro-N-nitrosoguanidine and thelike.

The strain No.14573 has the following morphological, cultural andphysiological characteristics.

The strain grew rather restrictedly on various culture media, and formedgrayish colonies. The strain produced pycnidial to stromatic conidiomataon the autoclaved leaf segments affixed on agar media, while it formedneither teleomorph nor anamorph on or in the media. The conidiomata wereconvex to discoid, dark brown to black, and formed ampulliform tolageniform conidiogenous cells on the lower cells of their inner walls.Conidia were hyaline, one-celled and cylindrical. On the basis ofcomparing the morphological characteristics with fungal taxonomiccriteria by von Arx (J. A. von Arx: The Genera of Fungi—Sporulating inPure Culture. 3rd ed., pp.315, J. Cramer, Vaduz, 1974) and by Sutton (B.C. Sutton: The Coelomycetes—Fungi Imperfecti with Pycnidia, Acervuli andStroma., pp.696, Commonwealth Mycological Institute, Kew, 1980), strainNo.14573 was considered to belong to the coelomycete genus ColeophomaHöhn. 1907 (Sphaeropsidales). Its mycological characteristics were asfollows.

Cultural characteristics on various agar media are summarized inTable 1. Culture on potato dextrose agar grew restrictedly, attaining1.5-2.5 cm in diameter two weeks later at 25° C. This colony surface wasplane to raised, felty to cottony, light gray to dark gray, andyellowish white at the margin. The reverse color was olive. Conidialstructures were not observed. Colonies on corn meal agar grew ratherrestrictedly, attaining 2.5-3.5 cm in diameter under the sameconditions. The surface was plane to centrally raised, dark gray andlustrous. At the colony center was felty to cottony and purplish gray todark purple. Mycelium near the margin was submerged and white. Thereverse was dark gray to dark green, and vellowish white at the margin.Conidial structures were not produced.

The morphological characteristics were determined from the cultures onthe sterile leaf segments affixed on a Miura's LCA plate (Miura, K. andM. Kudo: Trans. Mycol. Soc. Japan. 11:116-118, 1970). Conidiomata formedon the leaf segments alone. They were pycnidial to stromatic,superficial to semi-immersed, separate and dark brown to black. Theirshape was convex to discoid, sometimes papillate, non-ostiolate orindistinctly ostiolate, unilocular, flattened at the base, thin-walledat the upper part, 70-170 μm in diameter and 40-90 μm high. The lowercells of inner pycnidial walls were thick-walled, dark brown,irregularly shaped, and formed textura angularis. The inner cellsproduced directly conidiogenous cells, but they sometimes formedconidiophores. The conidiophores were hyaline, smooth, septate, simpleto sparingly branched, and 10-17×3.5-4.5 μm. The conidiogenous cellswere discrete, acrogenous or intercalary, hyaline, smooth, ampulliformto lageniform, sometimes cylindrical, and 5-11(-16)×2-4.5 μm. Conidiawere holoblastic, hyaline, smooth, one-celled, cylindrical, rounded atthe apical end, with a small projection at the base, and (11-)13-20×2-3μm. Paraphyses were often formed on or amongst conidiophores, and theirstructures were similar to sheaths covering with conidiogenous cells andconidia. They were hyaline, thin-walled, campanulate to cylindrical,collapsing at later stage, and 18-30(-35)×2.5-5 μm. Vegetative hyphaewere smooth, septate, brown and branched. The hyphal cells werecylindrical and 2-7 μm in width. Chlamydospores were not observed.

Strain No.14573 was able to grow at the temperature range from 3 to 30°C., with the growth optimum at 21 to 24° C. These temperature data weredetermined on potato dextrose agar (made by NISSUI).

According to the taxonomic criteria of the genus Coleophoma by Wu et al.(W. Wu, B. C. Sutton and A. C. Gange: Mycol. Res. 100: 943-947. 1996.),the strain No.14573 resembles Coleophoma empetri (Rostr.) Petrak 1929.There were few differences between above characteristics and thisspecies description: superficial and indistinctly ostiolate conidiomata.Added to these, it remains one question for the structures described asparaphyses. Much more observation was needed to determine the conidialontogeny of this genus. In conclusion, we identified the isolate as onestrain of Coleophoma empetri, and named it Coleophoma empetri No. 14573.

TABLE 1 Cultural characteristics of strain No. 14573. Media Culturalcharacteristics Malt extract agar* G: Very restrictedly, 1.0-2.0 cm S:Circular, plane, felty, formed no anamorph, brownish gray (5F2) tograyish brown (6F3) R: Olive (3F4) to olive brown (4F4), and olive(3E3-3E4) at the center Potato dextrose G: Restrictedly, 1.5-2.5 cm agar(Difco 0013) S: Circular, plane to raised, felty to cottony, formed noanamorph, light gray (1D1) to dark gray (1F1), yellowish white (3A2) atthe margin R: Olive (3D3-3F3) Czapek's solution G: Restrictedly, 1.5-2.5cm agar* S: Circular, submerged, thin, plane, formed no anamorph, olivebrown (4F4) R: Olive brown (4F4) Sabouraud dextrose G: Restrictedly,1.5-2.5 cm agar (Difco 0190) S: Circular, plane to centrally raised,felty, partly hygroscopic, sectoring, formed no anamorph, orange white(5A2) to grayish yellow (4C4), dark gray (1F1) at the center and sectorsR: Grayish yellow (4B4-4C3), and brownish gray (5F2) at the center andsectors Emerson Yp Ss agar G: Restrictedly, 2.0-3.0 cm (Difco 0739) S:Circular, plane, felty, formed no anamorph, gray (1F1) to olive gray(1F2), yellowish white (3A2) at the margin R: Greenish gray (25F2) todark green (25F3), and yellowish white (3A2) at the margin Corn mealagar G: Rather restrictedly, 2.5-3.5 cm (Difco 0386) S: Circular, planeto centrally raised, formed no anamorph, lustrous, dark gray (1F1):felty to cottony, purplish gray (14F2) to dark purple (14F3) at thecenter; submerged and white (1A1) at the margin R: Dark gray (1F1) todark green (25F3), yellowish white (3A2) at the margin MY20 agar* G:Very restrictedly, 1.0-2.0 cm S: Circular, plane, hygroscopic, lustrous,formed no anamorph, grayish yellow (4B3) R: Light yellow (4A4-4A5)Oatmeal agar G: Rather rapidly, 3.0-4.0 cm (Difco 0552) S: Circular,plane to centrally raised, felty to cottony, formed no anamorph, darkgray (1F1), and yellowish white (4A2) at the margin Abbreviation G:growth, measuring colony size in diameter, S: colony surface, R:reverse. *The compositions of malt extract agar, Czapek's solution agarand MY20 agar were based on JCM Catalogue of Strains (Nakase, T., 6thed., pp617., Japan Collection of Microorganisms, the Institute ofPhysical and Chemical Research, Saitama, 1995).

These characteristics were observed after 14 days of incubation at 25°C. The color descriptions were based on Methuen Handbook of Colour(Kornerup, A. and J. H. Wanscher, 3rd ed., pp.252., Methuen, London,1978).

Production of WF14573A and B

The compound, WF14573A and B can be prepared by culturing a WF14573Aand/or B-producing strain in a nutrient medium.

In general, WF14573A and B can be produced by culturing the WF14573Aand/or B-producing strain in a nutrient medium containing assimilablesources of carbon and nitrogen, preferably under aerobic conditions(e.g. shaking culture. submerged culture, etc.).

The preferred sources of carbon are carbohydrates such as sucrose,glucose, soluble starch and the like.

The preferred sources of nitrogen are cottonseed meal, soybean flour,yeast extract, peptone, gluten meal, corn steep liquor, dried yeast etc.as well as inorganic and organic nitrogen compounds such as ammoniumsalts (e.g. ammonium nitrate, ammonium sulfate, ammonium phosphate,etc.), urea, amino acid and the like.

The carbon and nitrogen sources need not be used in their pure form,because less pure materials which contain traces of growth factors andconsiderable quantities of mineral nutrients, are also suitable for use.Further, there may be added to the medium. Mineral salts such as calciumcarbonate, sodium or potassium phosphate magnesium salts and the like.If the culture medium is foamed remarkably, a defoaming agent such asliquid paraffin, higher alcohol, plant oil, mineral oil and siliconesmay be added.

Preferred production conditions of WF14573A and/or B in massive amountmay includes a submerged aerobic cultural condition.

Preferred production conditions of WF14573A and/or B in small amount mayinclude a shaking or surface culture in flask or bottle.

In case where the production is carried out in a large tank, it ispreferable to use the vegetative form of the organism for inoculation inthe production tank in order to avoid growth lag.

Agitation and aeration of the culture broth may be accomplished in avariety of ways. Agitation may be provided by a propeller or the similarmechanical agitation equipment, by revolving or shaking the fermentor,by various pumping equipment or by the passage of sterile air throughthe medium.

The fermentation is usually conducted at a temperature between 20° C.and 35° C., preferably about 25° C. for 50 to 100 hours, which may bevaried depending on the fermentation condition and scale.

Thus produced WF14573A and B can be recovered from the cultured broth byconventional means which are commonly used for the recovery of otherfermentation products such as antibiotics.

In general, most of the WF14573A and B produced are found in the culturefiltrate as well as in the cells. Accordingly, WF14573A and B can beisolated from the filtrate and the cells, which is obtained by filteringor centrifuging the broth, by a conventional method such asconcentration under reduced pressure, lyophilization, extraction with asolvent, pH adjustment, treatment with a resin (e.g. anion or cationexchange resin, non-ionic adsorption resin), treatment with an adsorbent(e.g. activated charcoal, silicic acid, silica gel, cellulose, alumina),crystallization, recrystallization and the like. The WF14573A and Bobtained in its free form may also be converted to its salts by treatingWF14573A and B with an inorganic or organic base such as sodium orpotassium hydroxide, ammonium hydroxide, ethanolamine and the like andwith an amino acid such as glycine, lysine, glutamic acid and the like.

The WF14573 and B have a strong antimicrobial activity againstpathogenic microorganisms, especially pathogenic fungi such aspathogenic yeast (e.g. Candida albicans etc.) and the like. Accordingly,the WF14573A and B and their pharmaceutically acceptable salt are usefulas an antimicrobial agent, especially antifungal agent which is used forthe treatment of infectious diseases in human beings and animals.

As examples for showing such pharmacological effects of WF14573A and B,some pharmacological test data are illustrated in the followings.

Test 1 (Antimicrobial Activity)

Antimicrobial activity of WF14573A or B was determined by a serial brothdilution method using 96-well microtiter plate in 100 μl of veastnitrogen base dextrose medium. The inoculum was adjusted to 1×10⁵ colonyforming units/ml. Candida albicans and Aspergillus fumigatus werecultured at 37° C. for 24 hours and Cyptococcus neoformans was culturedat 37° C. for 48 hours in 5% CO₂ incubator. After incubation, the growthinhibition of microorganism in each well was determined by microscopicobservation. The results were shown as MEC (minimum effectiveconcentration: μg/ml) value (Table 1).

TABLE 1 Antimicrobial activity of WF14573A and B. MEC (μg/ml)Microorganisms WF14573A WF14573B Candida albicans FP633 0.31 0.04Aspergillus fumigatus FP1305 0.08 0.02 Cryptococcus neoformans YC20350.0 50.0

The deacyl WF14573A or B or their salts can be prepared by deacylatingWF14573A or B or their salts, especially by deacylating WF14573A or B ortheir salts respectively, in the presence of cultured broth or itsprocessed material of a microoranism [such as a microorganism belongingto the genus Streptomyces, e.g. Streptomyces anulatus No. 4811 (FERMBP-5808)] which is capable of deacylating WF14573A or B to give deacylWF14573A or B.

The processed material of the cultured broth may include mycelia andcrude or purified deacylase preparations obtained from them.

The enzymatic reaction is carried out in a conventional manner, e.g.those described in the following working Examples.

The deacyl WF14573A or B can be converted to an acyl derivative thereofby a conventional acylation. The acyl derivative can be represented bythe above formula (I) wherein R² is acyl except palmitoyl.

The present antimicrobial agent comprising the WF14573 (I) orpharmaceutically acceptable salt thereof is useful as a therapeuticagent for infectious diseases in animals including human beings. Thepharmaceutically acceptable salt of the WF14573 (I) may include the saltas exemplified above.

The antimicrobial composition can be used in the form of pharmaceuticalpreparation, for example, in solid, semisolid or liquid form, whichcontains the WF14573 (I) or its salt in admixture with a pharmaceuticalorganic or inorganic carrier or excipient suitable for external, enteralor parenteral applications. The active ingredient may be compounded, forexample, with usual non-toxic, pharmaceutically acceptable carriers fortablets, pellets, capsules, suppositories, solutions, emulsions,suspensions, ointments and any other form suitable for use. Thepharmaceutically acceptable carriers are water, glucose, lactose, gumacacia, gelatin, mannitol, starch paste, magnesium trisilicate, talc,corn starch, keratin, colloidal silica, potato starch, urea and othercarriers suitable for use in manufacturing preparations and in addition,auxiliary, stabilizing, thickening and coloring agents and perfumes. Theantimicrobial compositions can also contain preserving or bacteriostaticagents thereby keeping the active ingredient in the desired preparationsstable in activity. The active object compound is contained in theantimicrobial composition in an amount sufficient to produce the desiredtherapeutic effect upon the bacterially infected process or condition.

For applying this composition to human patients, it is preferably toapply it in a form of intraveneous, intramuscular, oral or percutaneousadministration. While the dosage or therapeutically effective amount ofthe WF14573 (I) or pharmaceutically acceptable salts thereof variesdepending on the age, conditions of each individual patient to betreated, the preferred daily dosage of the WF14573 (I) can be selectedfrom the range of 0.1-100 mg/kg of the patient.

The following Examples are given for the purpose of illustrating thisinvention, but not limited thereto.

EXAMPLE 1 (1) Fermentation of Coleophoma empetri No. 14573 for theProduction of the WF14573A

An aqueous seed medium (30 ml) containing sucrose 4%, glucose 1%,soluble starch 2%, cottonseed meal 3%, soybean flour 1.5%, KH₂PO₄ 1%,CaCO₃ 0.2%, Adecanol LG-109 (defoaming agent, Asahi Denka Co., Ltd.)0.05%, and Silicone KM-70 (defoaming agent, Shin-Etsu Chemical Co.,Ltd.) 0.05% was placed in each of three 100-ml Erlenmeyer flasks and wassterilized at 120° C. for 30 minutes. A loopful of Coleophoma empetriNo. 14573, grown on YpSs agar at 25° C. for 2 weeks, was inoculated ineach of the seed flasks. The inoculated flasks were shaken on a rotaryshaker (220 rpm, 5.1 cm throw) at 25° C. for 5 days, and 8 ml of theseed culture were transferred to 160 ml of the same sterile seed mediumin the 500-ml Erlenmeyer flasks. The flasks were shaken on a rotaryshaker (220 rpm, 5.1 cm throw) at 25° C. for 2 days, and 640 ml (fourflasks) of the second seed culture were inoculated to 20 liters ofsterile production medium consisting of modified starch 5%, cottonseedmeal 2%, oat meal 0.5%, KH₂PO₄ 3.5%, Na₂HPO₄.12H₂O 2.63%, (NH₄)₂SO₄0.6%, L-isoleucine 0.5%, L-proline 0.5%, Adecanol LG-109 0.05%, andSilicone KM-70 0.05% in a 30-liter jar fermentor. Fermentation wascarried out at 25° C. for 5 days under aeration of 20 liters/minute andagitation of 250 rpm.

The production of the WF14573A in the fermentation broth was monitoredby HPLC analysis indicated below.

#analytical HPLC condition; column: YMC Pack ODS-AM 303, S-5 120A(250×4.6 mm I.D., YMC Co., Ltd.); mobile phase: 50% aqueous acetonitrilecontaining 0.1% TFA; flow rate: 1 ml/min. detection: UV at 210 nm;retention time: WF14573A 11.9 min.

(2) Isolation and Purification of the WF14573A

The culture broth (80 liters) was extracted with an equal volume ofacetone by stirring for 2 hours at room temperature. The mixture wasfiltered with an aid of diatomaceous earth. The filtrate was dilutedwith an equal volume of water and passed through a column (6 L) ofDIAION HP-20 (Mitsubishi Chemical Co., Ltd.) packed with water. Thecolumn was washed with 50% aqueous methanol (19 L) and then eluted withmethanol (34 L). The active fraction (0-20 L) was diluted with an equalvolume of water and passed through a column (4 L) of YMC-GEL (ODS-AM120-S50, YMC Co., Ltd.) packed with 50% aqueous methanol. The column waswashed with 50% (6.5 L) and 60% aqueous methanol (12L) and then elutedwith 70% aqueous methanol (15.9 L). The active fraction (5.8-15.9 L) wasconcentrated in vacuo to 5.3 L. One liter of this solution was dilutedwith an equal volume of water and passed through a column (1 L) ofYMC-GEL (ODS-AM 120-S50, YMC Co., Ltd.) packed with 20% aqueousmethanol. The column was washed with 40% aqueous acetonitrile containing0.5% NH₄H₂PO₄ (3 L) and eluted with 50% aqueous acetonitrile containing0.5% NH₄H₂PO₄ (2.9 L). The active fraction (1.05-1.35 L) was dilutedwith an equal volume of water and passed through a column (2 L) ofYMC-GEL (ODS-AM 120-S50, YMC Co., Ltd.) packed with 25% aqueousacetonitrile. The column was washed with 40% aqueous methanol and elutedwith 80% aqueous methanol. The active fraction was concentrated in vacuoto an aqueous solution and lyophilized to give 798 mg of crude WF14573A.This powder was dissolved in a small volume of water and furtherpurified by preparative HPLC, using YMC-packed column (ODS-AM SH-343-5AMS-5, 250×20 mm I.D., YMC Co., Ltd.) with 50% aqueous acetonitrilecontaining 0.5% NaH₂PO₄.2H₂O as a mobile phase. A flow rate was 9.9ml/minute. Fractions containing the WF14573A were collected. Theseactive fractions were diluted with an equal volume of water and passedthrough YMC-packed column (ODS-AM SH-343-5AM S-5, 250×20 mm I.D., YMCCo., Ltd.) equilibrated with 25% aqueous acetonitrile containing 0.25%NaH₂PO₄.2H₂O. The column was washed with 30% aqueous methanol and theneluted with 70% aqueous methanol at a flow rate of 9.9 ml/minute. Theeluate was concentrated in vacuo and lyophilized to give 121 mg ofWF14573A as white powder.

EXAMPLE 2 (1) Fermentation of Coleophoma empetri No. 14573 for theProduction of the WF14573B

An aqueous seed medium (30 ml) containing sucrose 4%, glucose 1%,soluble starch 2%, cottonseed meal 3%, soybean flour 1.5%, KH₂PO₄ 1%,CaCO₃ 0.2%, Adecanol LG-109 (defoaming agent, higher alcohol, AsahiDenka Co., Ltd.) 0.05%, and Silicone KM-70 (defoaming agent, Shin-EtsuChemical Co., Ltd.) 0.05% was placed in each of three 100-ml Erlenmeyerflasks and was sterilized at 120° C. for 30 minutes. A loopful ofColeophoma empetri No. 14573, grown on YpSs agar at 25° C. for 2 weeks,was inoculated in each of the seed flasks. The inoculated flasks wereshaken on a rotary shaker (220 rpm, 5.1 cm throw) at 25° C. for 5 days,and 8 ml of the seed culture was transferred to 160 ml of the samesterile seed medium in the 500-ml Erlenmeyer flasks. The flasks wereshaken on a rotarv shaker (220 rpm, 5.1 cm throw) at 25° C. for 2 days,and 640 ml (four flasks) of the second seed culture was inoculated to 20liters of sterile production medium consisting of sucrose 8%, driedyeast 4%, CaCO₃ 0.5%, Adecanol LG-109 0.05%, and Silicone KM-70 0.05%(pH 6.3 adjusted with 1N NaOH) in a 30-liter jar fermentor. Fermentationwas carried out at 25° C. for 5 days under aeration of 20 liters/minuteand agitation of 250 rpm.

The production of the WF14573B in the fermentation broth was monitoredby HPLC analysis indicated below.

#analytical HPLC condition; column: YMC Pack ODS-AM 303, S-5 120A(250×4.6 mm I.D., YMC Co., Ltd.); mobile phase: 50% aqueous acetonitrilecontaining 0.1% TFA; flow rate: 1 ml/min. detection: UV at 210 nm;retention time: WF14573B 13.2 min.

(2) Isolation and Purification of the WF14573B

The culture broth (40 liters) was extracted with an equal volume ofacetone by stirring for 2 hours at room temperature. The mixture wasfiltered with an aid of diatomaceous earth. The filtrate was dilutedwith an equal volume of water and passed through a column (3 L) ofDIAION HP-20 (Mitsubishi Chemical Co., Ltd.) packed with water. Thecolumn was washed with water (9 L) and 50% aqueous methanol (10 L) andthen eluted with methanol (29 L). The active fraction (0-20 L) wasdiluted with an equal volume of water and passed through a column (1 L)of YMC-GEL (ODS-AM 120-S50, YMC Co., Ltd.) packed with water. The columnwas washed with 60% (5 L) and 70% (2.8 L) aqueous methanol and theneluted with 80% aqueous methanol (2.8 L). The active fraction (0.8-2.8L) was diluted with an equal volume of water and passed through a column(1 L) of YMC-GEL (ODS-AM 120-S50, YMC Co., Ltd.) packed with water. Thecolumn was washed with 40% aqueous methanol (1 L) and eluted with 50%aqueous acetonitrile containing 0.5% NH₄H₂PO₄ (3.3 L). The activefraction (2.0-2.5 L) was diluted with an equal volume of water andpassed through a column (2 L) of YMC-GEL (ODS-AM 120-S50, YMC Co., Ltd.)packed with water. The column was washed with 40% aqueous methanol (6 L)and eluted with 80% aqueous methanol (4.35 L). The active fraction(2.5-4.35 L) was concentrated in vacuo to an aqueous solution andlyophilized to give 411 mgofcrude WF14573B. A part of this powder (120mg) was dissolved in a small volume of water and further purified bypreparative HPLC, using YMC-packed column (ODS-AM SH-343-5AM S-5, 250×20mm I.D., YMC Co., Ltd.) with 50% aqueous acetonitrile containing 0.5%NaH₂PO₄.2H₂O as a mobile phase. A flow rate was 9.9 ml/minute. Fractionscontaining the WF14573B were collected. These active fractions werediluted with an equal volume of water and passed through YMC-packedcolumn (ODS-AM SH-343-5AM S-5, 250×20 mm I.D., YMC Co., Ltd.)equilibrated with 25% aqueous acetonitrile containing 0.25%NaH₂PO₄.2H₂O. The column was washed with 30% aqueous methanol (240 ml)and then eluted with 80% aqueous methanol at a flow rate of 9.9ml/minute. The eluate was concentrated in vacuo and lyophilized to give70 mg of WF14573B as white powder.

EXAMPLE 3 Preparation of Deacyl WF14573A (1) Fermentation ofStreptomyces anulatus No. 4811 (FERM BP-5808)

A stock culture of Streptomyces anulatus No.4811 was prepared andmaintained on an agar slant. A loopful of the slant culture wasinoculated into 60 ml of sterilized seed medium consisting of maltose3%, dried yeast 1%, CaCO₃ 0.5% in a 225-ml Erlenmeyer flask. The flaskwas incubated at 30° C. for 3 days on a rotary shaker (220 rpm, 5.1cm-throw) and then inoculated (0.1%) into 60 ml of sterilized seedmedium consisting of maltose 3%, dried yeast 1%, CaCO₃ 0.5%, AdecanolLG-109 (Asahi Denka Co., Ltd.) 0.1% and Silicone KM-70 (Shin-EtsuChemical Co., Ltd.) 0.1% in each of three 225-ml Erlenmeyer flasks. Theflasks were incubated at 30° C. for 2 days on a rotary shaker (220 rpm,5.1 cm-throw).

The resultant seed culture was then inoculated (5%) into 60 ml ofsterilized production medium consisting of maltose 8%, soybean meal 2%,wheat germ 2%, potato protein 2%, CaCO₃ 0.5%, Adecanol LG-109 0.1% andSilicone KM-70 0.1% in each of fifty 225-ml Erlenmeyer flasks. Theflasks were incubated at 30° C. for 6 days on a rotary shaker (220 rpm,5.1 cm-throw). The vegetative mycelia were collected from the fermentedbroth by filtration and once washed with water. The washed mycelia wereused to obtain the deacyl WF14573A.

(2) Fermentation of Coleophoma empetri No. 14573 for the Production ofthe WF14573A

An aqueous seed medium (30 ml) containing sucrose 4%, glucose 1%,soluble starch 2%, cottonseed meal 3%, soybean flour 1.5%, KH₂PO₄ 1%,CaCO₃ 0.2%, Adecanol LG-109 (defoaming agent, Asahi Denka Co., Ltd.)0.05%, and Silicone KM-70 (defoaming agent, Shin-Etsu Chemical Co.,Ltd.) 0.05% was placed in each of three 100-ml Erlenmeyer flasks and wassterilized at 120° C. for 30 minutes. A loopful of Coleophoma empetriNo. 14573, grown on YpSs agar at 25° C. for 2 weeks, was inoculated ineach of the seed flasks. The inoculated flasks were shaken on a rotaryshaker (220 rpm, 5.1 cm throw) at 25° C. for 5 days, and 8 ml of theseed culture were transferred to 160 ml of the same sterile seed mediumin the 500-ml Erlenmeyer flasks. The flasks were shaken on a rotaryshaker (220 rpm, 5.1 cm throw) at 25° C. for 2 days, and 640 ml (fourflasks) of the second seed culture were inoculated to 20 liters ofsterile production medium consisting of modified starch 5%, cottonseedmeal 2%, oat meal 0.5%, KH2PO₄ 3.5%, Na₂HPO₄.12H₂O 2.63%, (NH₄)₂SO₄0.6%, L-isoleucine 0.5%, L-proline 0.5%, Adecanol LG-109 0.05%, andSilicone KM-70 0.05% in a 30-liter jar fermentor. Fermentation wascarried out at 25° C. for 5 days under aeration of 20 liters/minute andagitation of 250 rpm.

The production of the WF14573A in the fermentation broth was monitoredby HPLC analysis indicated below.

#analytical HPLC condition; column: YMC Pack ODS-AM 303, S-5 120A(250×4.6 mm I.D., YMC Co., Ltd.); mobile phase: 50% aqueous acetonitrilecontaining 0.1% TFA; flow rate: 1 m/min. detection: UV at 210 nm;retention time: WF14573A 11.9 min.

(3) Preparation of the Crude WF14573A

The culture broth (80 liters) was extracted with an equal volume ofacetone by stirring for 2 hours at room temperature. The mixture wasfiltered with an aid of diatomaceous earth. The filtrate was dilutedwith an equal volume of water and passed through a column (6 L) ofDIAION HP-20 (Mitsubishi Chemical Co., Ltd.) packed with water. Thecolumn was washed with 50% aqueous methanol (19 L) and then eluted withmethanol (34 L). The active fraction (0-20 L) was diluted with an equalvolume of water and passed through a column (4 L) of YMC-GEL (ODS-AM120-S50, YMC Co., Ltd.) packed with 50% aqueous methanol. The column waswashed with 50% (6.5 L) and 60% (12 L) aqueous methanol and then elutedwith 70% aqueous methanol (15.9 L). The active fraction (5.8-15.9 L) wasconcentrated in vacuo to 5.3 L. A part (4.3 L) of this solution was usedto obtain the deacyl WF14573A.

(4) Deacylation of WF14573A

In the 30-liter jar fermentor, 900 ml of 1M sodium phosphate buffer(pH5.8) and 400 g of the vegetative mycelia of Streptomyces anulatusNo.4811 were added to this solution. The reaction mixture was filled upto 20 L with water. The reaction was carried out at 50° C. with stirringfor 2 hours. Decrease of the WF14573A was monitored by analytical HPLCindicated before and increase of the deacyl WF14573A was monitored byanalytical HPLC indicated below.

#analytical HPLC condition; column: YMC Pack ODS-AM 303, S-5 120A(250×4.6 mm I.D., YMC Co., Ltd.); mobile phase: 6.7% aqueousacetonitrile containing 0.1% TFA; flow rate: 1 m/min. detection: UV at210 nm; retention time: deacyl WF14573A 8.8 min.

(5) Isolation of the Deacyl WF14573A

The reaction mixture described above was filtered with an aid ofdiatomaceous earth. The mycelial cake was discarded. The filtrate thusobtained was concentrated in vacuo to 11 L, and passed through a column(1 L) of SEPABEADS SP-207 (Mitsubishi Chemical Co., Ltd.) packed withwater. The column was washed with water (3 L) and then eluted with 50%aqueous methanol (3 L). The active fraction (0-2 L) was concentrated invacuo to an aqueous solution. This solution was passed through a column(2 L) of YMC GEL (ODS-AM 120-S50, YMC Co., Ltd.) packed with water. Thecolumn was washed with water and 5% aqueous methanol containing 0.5%NaH₂PO₄.2H₂O (5 L) and 7% aqueous methanol containing 0.5% NaH₂PO₄.2H₂O(3.8 L) and eluted with 10% aqueous methanol containing 0.5%NaH₂PO₄.2H₂O (13.6 L). The active fraction (1.9-11.6 L) was concentratedin vacuo to 3 L. One liter of this solution was passed through a column(2 L) of YMC GEL (ODS-AM 120-S50, YMC Co., Ltd.) packed with water. Thecolumn was washed with water and 7% aqueous methanol containing 0.5%NaH₂PO₄.2H₂O (4 L) and eluted with 11% aqueous methanol containing 0.5%NaH₂PO₄.2H₂O (9.2 L). The active fracrion (5.3-6.8 L) was diluted withan equal volume of water and passed through a column (2 L) of YMC GEL(ODS-AM 120-S50) packed with water. The column was washed with water andeluted with 20% aqueous methanol. The eluate was concentrated in vacuoand lyophilized to give 820 mg of the deacyl WF14573A as a white powder.One hundred twenty milligrams of this powder were dissolved in a smallvolume of water and further purified by preparative HPLC, usingYMC-packed column (ODS-AM SH-343-5AM S-5, 250×20 mm I.D., YMC Co., Ltd.)with 11% aqueous methanol containing 0.5% NaH₂PO₄.2H₂O as a mobile phaseand a flow rate of 9.9 ml/minute. Active fraction was diluted with anequal volume of water and passed through YMC-packed column (ODS-AMSH-343-5AM S-5, 250×20 mm I.D., YMC Co., Ltd.) equilibrated with water.The column was washed with water (240 ml) and then eluted with 20%aqueous methanol at a flow rate of 9.9 ml/minute. The eluate wasconcentrated in vacuo and lyophilized to give 67.7 mg of deacyl WF14573Aas white powder.

¹H NMR spectrum (500 MHz, D2O) δ (ppm): 7.29 (1H, d, 8), 6.86 (1H, d,2), 6.77 (1H, dd, 8, 2), 5.41 (1H, d, 3), 5.06 (1H, d, 3), 4.94 (1H, d,6), 4.73 (1H, m), 4.66˜4.60 (2H, m), 4.54˜4.46 (3H, m),4.40 (1H, m),4.24˜4.19 (2H, m), 4.14˜4.07 (2H, m), 3.92 (1H, m), 3.84˜3.77 (2H, m),2.78˜2.67 (2H, m), 2.55 (1H, m), 2.46˜2.32 (4H, m), 2.15 (1H, m),2.08˜1.99 (2H, m),1.24 (3H, d, 6). ¹³C spectrum (125 MHz, D2O) δ (ppm):176.2 (s), 174.6 (s), 172.24 (s), 172.24 (s), 172.24 (s), 169.7 (s),169.0 (s), 148.5 (s), 138.2 (s), 137.2 (s), 123.5 (d), 122.1 (d), 118.6(d), 75.8 (d), 73.1 (d), 72.3 (d), 70.7 (d), 70.6 (d),69.3 (d), 68.0(d). 67.3 (d), 61.7 (d), 58.0 (d), 57.4 (d), 56.3 (t), 54.8 (d), 53.1(d), 46.4 (t), 39.8 (t), 39.7 (t), 37.6 (t), 33.1 (t), 31.6 (t), 19.2(q).

EXAMPLE 4 Preparation of Deacyl WF14573B (1) Fermentation ofStreptomyces anulatus No. 4811 (FERM BP-5808).

A stock culture of Streptomyces anulatus No.4811 was prepared andmaintained on agar slant. A loopful of the slant culture was inoculatedinto 60 ml of sterilized seed medium consisting of maltose 3%, driedyeast 1%, CaCO₃ 0.5% in a 225-ml Erlenmeyer flask. The flask wasincubated at 30° C. for 3 days on a rotary shaker (220 rpm, 5.1cm-throw) and then inoculated (0.1%) into 60 ml of sterilized seedmedium consisting of maltose 3%, dried yeast 1%, CaCO₃ 0.5%, AdekanolLG-109 (Asahi Denka Co., Ltd.) 0.1% and Silicone KM-70 (Shin-Etsuchemical Co., Ltd.) 0.1% in each of three 225-ml Erlenmeyer flasks. Theflasks were incubated at 30° C. for 2 days on a rotary shaker (220 rpm,5.1 cm-throw).

The resultant seed culture was then inoculated (5%) into 60 ml ofsterilized production medium consisting of maltose 8%, soybean meal 2%,wheat germ 2%, potato protein 2%, CaCO₃ 0.5%, Adekanol LG-109 0.1% andSilicone KM-70 0.1% in each of fifty 225-ml Erlenmeyer flasks. Theflasks were incubated at 30° C. for 6 days on a rotary shaker (220 rpm,5.1 cm-throw). The vegetative mycelium was collected from the fermentedbroth by filtration and once washed with water. The washed mycelium wasused to obtain the deacyl WF14573B.

(2) Fermentation of Coleophoma empetri No. 14573 for the Production ofthe WF14573B

A loopful of the slant culture (C. empetri No. 14573) was inoculatedinto 60 ml of sterilized seed medium consisting of sucrose 4%, glucose1%, soluble starch 2%, cotton seed flour 3%, soybean powder 1.5%. KH₂PO₄1%, CaCO₃ 0.2%, Adekanol LG-109 0.05% and Silicone KM-70 0.05% in eachof three 225-ml Erlenmeyer flasks. The flasks were incubated at 25° C.for 5 days on a rotary shaker (220 rpm, 5.1 cm-throw) and theninoculated (5%) into 160 ml of the same sterilized seed medium in eachof twenty 500-ml Erlenmever flasks. The flasks were incubated at 25° C.for 2 davs on a rotary shaker (220 rpm, 5.1 cm-throw).

The resultant seed culture was inoculated (3%) into 20 liters ofsterilized production medium in each of five 30-liter jar fermenters.The production medium was composed of sucrose 8%, dried veast 4%, CaCO₃0.5%, Adekanol LG-109 0.05% and Silicone KM-70 0.05%. The pH wasadjusted to 6.3 prior to sterilization. The fermentation was carried outat 25° C. for 5 days under aeration of 20 liters/minute and agitation of250 rpm.

The production of the WF14573B substance in the fermentation broth wasmonitored by HPLC analysis indicated below.

#analytical HPLC condition; column: YMC Pack ODS-AM 303, S-5 120A(250×4.6 mm I.D., YMC Co., Ltd.); mobile phase: 50% aqueous acetonitrilecontaining 0.1% TFA; flow rate: 1 m/min. detection: UV at 210 nm;retention time: WF14573B 13.2 min.

(3) Preparation of the Crude WF14573B Substance

After the culture was completed, an equal volume of acetone was added tothe cultured broth. The mixture was allowed to stand for about one hourwith stirring at room temperature. The resultant mixture was filtratedwith an aid of diatomaceous earth. The filtrate was diluted with anequal volume of water and passed through a column (7 L) of DIAION HP-20(Mitsubishi Chemical Co., Ltd.) packed with water. The column was washedwith water (18 L) and 50% aqueous methanol (20 L) and then eluted withmethanol (35 L). The eluate was concentrated in vacuo to an aqueoussolution. This solution was passed through a column (2 L) of YMC-GEL(ODS-AM 120-S50, YMC Co., LTD.) packed with water. The column was washedwith water (6 L) and 50% aqueous methanol (4 L) and then eluted with 80%aqueous methanol (8 L). The elute was concentrated in vacuo to anaqueous solution (600ml).

(4) Deacylation of WF14573B

Thirty ml of 1M Na-phosphate buffer (pH5.8) and 60 g of the vegetativemycelium of Streptomyces anulatus No. 4811 (FERM BP-5808) were added tothis solution. The reaction was carried out at 50° C. with stirring for1 hour. Decrease of the WF14573B was monitored by analytical HPLCindicated before and increase of the deacyl WF14573B was monitored byanalytical HPLC indicated below.

From 663 mg of the WF14573B, 362 mg of the deacyl WF14573B was formed inthe reaction mixture.

#analytical HPLC condition; column: YMC Pack ODS-AM 303, S-5 120A(250×4.6 mm I.D., YMC Co., Ltd.); mobile phase: 10% aqueous methanolcontaining 0.1% TFA; flow rate: 1 m/min. detection: UV at 210 nm;retention time: deacyl WF14573B 11.4 min.

(5) Isolation of the Deacyl WF14573B

The reaction mixture described above was filtrated with an aid ofdiatomaceous earth. The mycelial cake was discarded. The filtrate thusobtained was passed through a column (150 ml) of SEPABEADS SP-207(Mitsubishi Chemical Co., Ltd.) packed with water. The column was washedwith water (450 ml) and then eluted with 50% aqueous methanol (450 ml).The eluate was concentrated in vacuo to an aqueous solution (100 ml).This solution was passed through a column (180 ml) of YMC GEL (ODS-AM120-S50, YMC Co., Ltd.) packed with water. The column was washed withwater and 5% aqueous methanol containing 0.5% NaH₂PO₄.2H₂O (600 ml) andeluted with 7.5 and 10% aqueous methanol containing 0.5% NaH₂PO₄.2H₂Orespectively. The elution was monitored by analytical HPLC indicatedbefore. The portion corresponding to the deacyl WF14573B wasconcentrated in vacuo to give residual water. This residue was passedthrough a column (180 ml) of YMC GEL (ODS-AM 120-S50) packed with water.The column was washed with water and 5% aqueous methanol (600 ml) andeluted with 7.5 and 10% aqueous methanol. The eluate was concentrated invacuo and lyophilized to give 228 mg of the deacyl WF14573B as whitepowder. Eighty mg of this powder was dissolved in a small volume ofwater and further purified by preparative HPLC, using YMC-packed column(ODS-AM SH-343-5AM S-5, 250×20 mm I.D., YMC Co., Ltd.) with 12% aqueousmethanol containing 0.5% NaH₂PO₄.2H₂O as a mobile phase and a flow rateof 9.9 ml/minute. Fractions containing the deacyl WF14573B werecollected and concentrated in vacuo to give residual water. This residuewas passed through YMC-packed column (ODS-AM SH-343-5AM S-5, 250×20 mmI.D., YMC Co., Ltd.) equilibrated with water. The column was washed withwater (240 ml) and then eluted with 20% aqueous methanol at a flow rateof 9.9 ml/minute. The eluate was concentrated in vacuo and lyophilizedto give 41 mg of deacyl WF14573B as white powder.

¹H NMR spectrum (500 MHz, D2O) δ (ppm): 7.28 (1H, d, 8), 6.85 (1H, d,2), 6.76 (1H, dd, 8, 2), 5.40 (1H, d, 3), 5.04 (1H, d, 3), 4.92 (1H, d,6), 4.72 (1H, m), 4.66-4.60 (2H, m), 4.54˜4.47 (2H, m), 4.44˜4.37 (2H,m), 4.29 (1H, d, 5), 4.22 (1H, m), 4.10˜4.06 (2H, m), 3.93˜3.89 (2H, m),3.42 (1H, m), 2.77˜2.66 (2H, m), 2.60˜2.50 (2H, m), 2.46˜2.35 (3H, m),2.14 (1H, m), 2.02 (1H, m), 1.24 (3H, d, 7), 1.02 (3H, d, 7). ¹³C NMRspectrum (125 MHz, D2O) δ (ppm): 178.4 (s), 176.7 (s), 174.49 (s),174.45 (s), 174.40 (s), 171.9 (s), 171.4 (s), 150.7 (s), 140.4 (s),139.4 (s), 125.7 (d), 124.3 (d), 120.8 (d), 78.1 (d), 76.9 (d), 74.5(d), 72.95 (d), 72.95 (d), 71.4 (d), 69.45 (d), 69.40 (d), 63.9 (d),60.2 (d), 59.6 (d), 58.5 (t), 57.0 (d), 55.4 (d), 54.9 (t), 42.01 (t),41.94 (t), 39.85 (d), 39.85 (t), 33.7 (t), 21.4 (q), 13.4 (q).

What is claimed is:
 1. A compound of formula (I):

wherein R¹ is hydrogen or methyl and R² is hydrogen or acyl, or a saltthereof.
 2. The compound of claim 1, wherein R¹ is hydrogen and R² ispalmitoyl.
 3. The compound of claim 1, wherein R¹ is methyl and R² ispalmitoyl.
 4. The compound of claim 1, wherein R¹ is hydrogen and R² ishydrogen.
 5. The compound of claim 1, wherein R¹ is methyl and R² ishydrogen.
 6. A salt of the compound of claim
 1. 7. A compositioncomprising the compound of claim 1 and optionally, one or more auxiliaryagent(s), stabilizing agent(s), thickening agent(s), coloring agent(s),perfume(s), preservative(s) or bacteriostatic agent(s).
 8. Thecomposition of claim 7 in combination with a pharmaceutically acceptablecarrier or excipient.
 9. The composition of claim 7, further comprisingone or more auxiliary agent(s), stabilizing agent(s), thickeningagent(s), coloring agent(s), or perfume(s).
 10. The composition of claim7, further comprising one or more preservative(s) or bacteriostaticagent(s).
 11. The composition of claim 7 in the form of a tablet,pellet, or capsule.
 12. The composition of claim 7 in the form of asolution, emulsion or suspension.
 13. The composition of claim 7 in theform of a suppository or ointment.
 14. A method for inhibiting thegrowth of a yeast or fungus comprising contacting a yeast or fungus withan amount of the compound of claim 1 effective to inhibit the growth ofsaid yeast or fungus.
 15. The method of claim 14, wherein said yeast orfungus is Candida.
 16. The method of claim 14, wherein said yeast orfungus is Aspergillus.
 17. The method of claim 14, wherein said yeast orfungus is Cryptococcus.
 18. A process for producing a compound offormula (I):

wherein R¹ is hydrogen or methyl and R² is hydrogen or palmitoyl, or asalt thereof, comprising: (a) culturing a microorganism that producesthe compound of formula (I) or a salt thereof, and (b) recovering thecompound of formula (I) or a salt thereof.
 19. The process of claim 18,wherein said microorganism is Coleophoma.
 20. The process of claim 18,wherein said microorganism is Coleophoma empetri.
 21. The process ofclaim 18, wherein said microorganism is Coleophoma empetri No. 14573.22. A process for producing the compound of formula (I):

wherein R¹ is hydrogen or methyl and R² is hydrogen, or a salt thereofcomprising: deacylating a compound of formula (I) by using deacylase,wherein R² is palmitoyl and recovering the compound of formula (I)wherein R¹ is hydrogen or methyl and R² is hydrogen, or a salt thereof.23. The process of claim 22, comprising contacting said compound offormula (I), wherein R² is palmitoyl with a microorganism to deacylatesaid compound.
 24. The process of claim 22, comprising contacting saidcompound of formula (I), wherein R² is palmitoyl, with Streptomyces todeacylate said compound.
 25. The method of claim 14, comprisingintravenously administering to a subject in need thereof an amount ofsaid compound effective to inhibit the growth of said yeast or fungus.26. The method of claim 14, comprising intramuscularly administering toa subject in need thereof an amount of said compound effective toinhibit the growth of said yeast or fungus.
 27. The method of claim 14,comprising orally administering to a subject in need thereof an amountof said compound effective to inhibit the growth of said yeast orfungus.
 28. The method of claim 14, comprising percutaneouslyadministering to a subject in need thereof an amount of said compoundeffective to inhibit the growth of said yeast or fungus.